Quantitative detection of micrometastases in the lymph nodes of gastric cancer patients with real-time RT-PCR: a comparative study with immunohistochemistry

Histologic examination lacks the sensitivity to detect micrometastases in gastric cancer lymph nodes. In the present study, we applied a real-time RT-PCR approach to the quantitative detection of micrometastases in gastric cancer lymph nodes and compared diagnostic power with routine histology and immunohistochemistry. We studied 392 lymph nodes from 21 gastric cancer patients who underwent curative surgery. Real-time quantitative RT-PCR was performed on a LightCycler instrument using a hybridization probe for carcinoembryonic antigen (CEA) and cytokeratin-20 (CK20) as marker genes. Immunohistochemistry with antibodies to wide-keratin was also performed in the lymph nodes to compare the sensitivity and specificity. Median (average) values of CEA mRNA in lymph nodes in patients with histology(+), immunohistochemistry(+)/histology(-), immunohistochemistry(-)/histology(-) and negative control results were 4600 (16000), 200 (400), 0 (9.8) and 0 (0.6), respectively. There were some false-negative results with simple CEA and CK20 real-time RT-PCR due to the presence of low gene-expressing gastric cancers as revealed by CEA and CK20 immunohistochemistry. CEA in combination with CK20 (duplex) real-time RT-PCR partially covered this weakness. Consequently, all 71 histology(+) lymph nodes were positive for duplex real-time RT-PCR as well as wide-keratin immunohistochemistry. Positivity rates by histology, wide-keratin immunohistochemistry and duplex real-time RT-PCR were 18.0% (71/392), 20.9% (82/392) and 25.8% (101/392), respectively. In 2 of 8 patients with pT1N0, positive lymph nodes were observed by real-time RT-PCR but not by immunohistochemistry. These results indicate that duplex quantitative real-time RT-PCR is the most sensitive method for detecting micrometastases and useful for evaluating the prognostic significance of lymph node micrometastasis in gastric cancer patients.     

Comparative immunocytochemical assessment of isolated carcinoma cells in lymph nodes and bone marrow of patients with clinically localized prostate cancer

After radical prostatectomy for clinically localized prostate cancer, biochemical progression is seen in up to 40% of the patients due to persistent local and/or systemic remnants. Isolated disseminated carcinoma cells, undetectable by current staging methods, are of special interest as potential precursors of subsequent overt metastases. In the present study, immunohistochemistry (IHC) was performed to evaluate simultaneously the frequency of occult carcinoma cells in both lymph nodes (LNs) and bone marrow (BM) obtained from the iliac crests of 45 prostate cancer patients with untreated stage T(1-3) pN0 M0 prostatic carcinoma. IHC using monoclonal antibodies (MAbs) against epithelial cytokeratins was performed on 521 paraffin-embedded LNs histopathologically classified as tumorfree (pN0), as well as on BM cytospin preparations. To confirm the prostatic origin of positive cells in LNs, additional IHCs for prostate-specific antigen (PSA) and epithelial glycoproteins were performed. In total, isolated tumor cells in LNs and/or BM were detected in 17 of the 45 patients. Parameters such as tumor stage, grade and volume of the primary tumor as well as blood serum PSA levels could not detect patients harboring disseminated single tumor cells in LNs or BM. Following a median observation time of 24.9 months, no significant correlation between IHC positivity and PSA progression as a measure of early relapse was observed. Although the overall incidence of occult tumor cell spread corresponds to similar incidence of relapses after radical prostatectomy as reported by others, the fate of these cells needs to be evaluated in longer follow-up studies.     

A study of image-guided intensity-modulated radiotherapy with fiducials for localized prostate cancer including pelvic lymph nodes

PURPOSE: To study the impact on nodal coverage and dose to fixed organs at risk when using daily fiducial localization of the prostate to deliver intensity-modulated radiotherapy (IMRT). METHODS AND MATERIALS: Five patients with prostate cancer in whom prostate and pelvic nodes were irradiated with IMRT were studied. Dose was prescribed such that 95% of the prostate planning target volume (PTV) and 90% of the nodal PTV were covered. Random and systematic prostate displacements in the anterior-posterior, superior-inferior, and left-right directions were simulated to shift the original isocenter of the IMRT plan. The composite dose during the course of treatment was calculated. RESULTS: Compared with a static setup, simulating random shifts reduced dose by less than 1.5% for nodal hotspot (i.e., dose to 1 cm(3)), by less than 1% for the 90% nodal PTV coverage, and by less than 0.5% for the nodal mean dose. Bowel and femoral head hotspots were reduced by less than 1.5% and 2%, respectively. A 10-mm systematic offset reduced nodal coverage by up to 10%. CONCLUSION: The use of prostate fiducials for daily localization during IMRT treatment results in negligible changes in dose coverage of pelvic nodes or normal tissue sparing in the absence of a significant systematic offset. This offers a simple and practical solution to the problem of image-guided radiotherapy for prostate cancer when including pelvic nodes.     

Correlation between side of palpable tumor and side of pelvic lymph node metastasis in clinically localized prostate cancer.

Of 411 patients with palpable but clinically localized (Stages B or C) adenocarcinoma of the prostate, 100 (24.3%) were found at complete bilateral pelvic lymphadenectomy to have one or more lymph nodes positive for metastasis. These patients were divided into five subgroups on the basis of the location of the palpable tumor at digital rectal examination: left side only, left predominantly, both sides, right side predominantly, or right side only. Among 35 patients with positive nodes and a palpable tumor limited to one side of the prostate (clinically unilobar), metastases were found in the ipsilateral pelvic lymph nodes in 29 (83%). Only 6 (17%) of the patients had contralateral metastasis alone. A unilateral pelvic lymphadenectomy (ipsilateral to the side of the largest palpable tumor, or on either side if the tumor were bilateral) would have detected 80% of the patients with positive lymph nodes, with a positive predictive value of 100% and a negative predictive value of 94%. Lymph node metastases in patients with clinically localized palpable prostate cancer are most likely to be found on the same side as the palpable tumor and are considerably less likely to be found on the contralateral side alone. If frozen section examination of lymph nodes or laparoscopic lymph node dissection is planned before definitive therapy for prostate cancer, the pelvic lymph nodes ipsilateral to the side of the palpable tumor should be removed first.     

Use of preoperative plasma endoglin for prediction of lymph node metastasis in patients with clinically localized prostate cancer.

PURPOSE: Current predictive tools and imaging modalities are not accurate enough to preoperatively diagnose lymph node metastases in patients with prostate cancer. The aim of the study was to evaluate whether preoperative plasma endoglin improves the prediction of lymph node metastases in patients with clinically localized prostate cancer. EXPERIMENTAL DESIGN: Endoglin levels were measured using a commercially available ELISA assay in banked plasma from 425 patients treated with radical prostatectomy and bilateral lymphadenectomy for clinically localized prostatic adenocarcinoma at two university hospitals between July 1994 and November 1997. Logistic regression analyses were undertaken to evaluate whether endoglin improves the accuracy of a standard preoperative model for prediction of lymph node metastasis and to build a predictive nomogram. RESULTS: Preoperative plasma endoglin levels were higher in patients with higher preoperative total serum prostate-specific antigen (PSA; Spearman correlation coefficient 0.296, P < 0.001), positive surgical margins (P = 0.03), higher pathologic Gleason sum (P = 0.04), and lymph node metastasis (P < 0.001). In a preoperative multivariable logistic regression analysis that included PSA and clinical stage, only preoperative endoglin (odds ratio, 1.17; 95% confidence interval, 1.09-1.26; P < 0.001) and biopsy Gleason sum (odds ratio, 18.57; 95% confidence interval, 1.08-318.36; P = 0.04) were associated with metastasis to lymph nodes. The addition of endoglin to a standard preoperative model (including PSA, clinical stage, and biopsy Gleason sum) significantly improved its accuracy for prediction of lymph node metastasis from 89.4% to 97.8% (P < 0.001). CONCLUSIONS: Preoperative plasma endoglin improves the accuracy for prediction of pelvic lymph node metastasis in patients treated with radical prostatectomy for clinically localized prostate cancer by a statistically and clinically significant margin.     

Quantification of micrometastases in lymph nodes of colorectal cancer using real-time fluorescence polymerase chain reaction

The expression of carcinoembryonic antigen (CEA) mRNA was assessed in 102 lymph nodes (LNs) obtained from seven colorectal cancer patients by both the conventional non-quantitative RT-PCR and quantitative RT-PCR. The number of CEA-expressing cells was calculated compared with CEA-expressing MKN-45 cell line as a standard control. Using the quantitative RT-PCR, the relative number of CEA-expressing cells ranged between 1.3x103 and 5.7x106 in 16 histologically positive LNs and between 2.3x101 and 8.1x105 in 10 histologically negative and RT-PCR positive LNs. In both histologically and RT-PCR negative LNs, the relative cell number was <4.0x102. Our results demonstrated that quantifying the amount of metastasis might enhance the reliability of RT-PCR detection assay as a diagnostic tool for the detection of cancer micrometastases.     

Lymphography in clinically localized prostate cancer

Lymphography demonstrates the size, position, and internal architecture of the external iliac, common iliac, para-aortic, and paracaval lymph nodes. Importantly, the "surgical obturator" nodes are also routinely opacified because they are part of the external iliac chain. Analysis of the internal architecture permits detection of metastases in nodes of normal size, an advantage over cross-sectional imaging techniques. In a prospective study of 89 unselected, previously untreated patients with carcinoma limited to the prostate or periprostatic bed, lymphography was compared with histology of lymph nodes removed at surgical staging. The sensitivity was 53% (17 of 32), specificity 93% (53 of 57), accuracy 79% (70 of 89), and positive and negative predictive values were 81% (17 of 21) and 78% (53 of 68), respectively.     

Intensity-modulated radiotherapy for the treatment of pelvic lymph nodes in prostate cancer

Evaluation of: Wang-Chesebro A, Xia P, Coleman J, Akazawa C, Roach M 3rd: Intensity-modulated radiotherapy improves lymph node coverage and dose to critical structures compared with 3D conformal radiation therapy in clinically localized prostate cancer. Int. J. Radiat. Oncol. Biol. Phys. 66, 654-662 (2006). A large randomized Phase III trial (RTOG 94-13) demonstrated improved progression-free survival for the irradiation of the pelvic lymphatics compared with treatment of the prostate only in patients with a high risk of lymph node involvement. Recent studies have indicated that the conventional target volume might miss substantial parts of the lymphatic drainage of the prostate. This retrospective planning study compared conventional, 3D-conformal and intensity-modulated radiotherapy (IMRT) for the treatment of pelvic lymph nodes. Field-shaping based on bony landmarks was shown to result in inadequate target coverage compared with 3D-conformal and IMRT planning. Regarding sparing of rectum, bladder, small bowl and penile bulb, the IMRT plans were highly superior. In summary, IMRT may result in increased rates of regional control with simultaneously decreased rates of toxicity. Integration of functional imaging into treatment planning and image guidance during treatment is expected to further improve the therapeutic ratio.     

Keratin 19 mRNA measurement to detect micrometastases in lymph nodes in breast cancer patients.

We have used polymerase chain reaction (PCR) to measure keratin 19 mRNA in order to detect breast cancer cells invading axillary lymph nodes. In a consecutive series of 125 patients with primary breast cancer, 75 patients had no evidence of lymph node involvement by conventional histology. A total of 530 lymph nodes from these patients were examined and 106 (20%) gave a keratin 19 product detectable by Southern hybridisation. This correlated with primary tumour size (P<0.001). These 106 nodes came from 23 patients. Thus, using this technique, 23/75 (30.6%) patients were found to have evidence of lymph node involvement who would otherwise have been designated lymph node negative.     

RT-PCR法检测肺癌患者区域性淋巴结微转移的临床病理相关性分析

目的：分析RT－PCR法检测区域性淋巴结中肺癌微转移的临床病理相关性。方法：区域性淋巴结共261枚，取自40例接受手术治疗的原发性肺癌患者，将每枚淋巴结均分为两等份，分别进行病理学检测和细胞骨架角蛋白19（CK19）基因的表达分析。结果：18例患者同时被病理检查和RT－PCR法证实存在淋巴结转移，另22例病理学检查未检淋巴结转移，但RT－CR法检测到其中6例存在淋巴结肺癌微转移。40例患者中，RT－PCR法显示淋巴结转移与肿瘤大小，癌肿侵犯血管，肿瘤分化等级和肿瘤的P－TNM分期有密切关系（P＜0．05）。22例病理检查未发现淋巴结转移的患者中，淋巴结微转移与肿瘤大小和肿瘤的P－TNM分期有密切关系（P＜0．05）。普通病理检查则显示淋巴结转移与否与患者的各临床病理指标之间无明显关系。结论：RT－PCR法在检测淋巴结转移方面较普通病理检查优越，它能准确地检测到存在于淋巴结中的肿瘤微小转移灶，有利于筛筛选早期亚临床转移的人群和揭示肿瘤转移的内在规律。     

External validation of the Briganti 2019 nomogram to identify candidates for extended pelvic lymph node dissection among patients with high-risk clinically localized prostate cancer

International Journal of Clinical Oncology - We aimed to establish an external validation of the Briganti 2019 nomogram in a Japanese cohort to preoperatively evaluate the probability of lymph node...     

Patterns of lateral pelvic lymph node metastases and micrometastases for patients with lower rectal cancer.

AIMS: We aimed at investigating the patterns of lymph node metastases and micrometastases in regions of lateral pelvic area, examining circumferential margin involvement and clarifying their prognostic significance. METHODS: Large tissue slice and tissue array were adopted in the study of 67 patients with AJCC stages I-III lower rectal cancer who underwent total mesorectal excision with systematic lateral pelvic dissection. The outcomes were followed. RESULTS: Altogether, 726 lateral lymph nodes were examined, with 32 and 38 were involved by tumor metastases and micrometastases, respectively. Fifty-eight (82.9%) of the involved lymph nodes were smaller than 5mm. Status of lateral nodes was related to that of mesorectal ones. Middle rectal root (45.5%), internal iliac (31.8%) and obturator (22.7%) regions were more likely to be involved by metastases. Patients with lateral metastases, similar to the group with micrometastases, suffered more recurrence and poorer survival when compared with the ones without metastases. The occurrence of circumferential margin involvement suggested poor prognosis and was related to lateral node status. CONCLUSIONS: In lateral pelvic area, the majority of lymph nodes harboring tumor were small and could easily be neglected by conventional examination. Incidence of lateral metastases differed among regions, thus more attention should be given to the clearance of the highly occurred areas. More extensive range of dissection and/or adjuvant therapy was recommended for patients with lateral node metastases, micrometastases and circumferential margin involvement, since they predisposed poor prognosis.     

Histologic detection and clinical implications of micrometastases in axillary sentinel lymph nodes for patients with breast carcinoma.

BACKGROUND: Sentinel lymph node (SLN) biopsy is used increasingly in patients with clinically lymph node negative, early-stage breast carcinoma, because it can spare axillary dissection when the sentinel lymph nodes are negative. The question arises, however, whether complete axillary lymph node dissection (ALND) also is necessary in patients with only micrometastases (< or = 2 mm in greatest dimension) in axillary SLNs. The authors carried out the current study to ascertain the risk of non-SLN axillary metastases in such patients and to assess the detection rate of SLN micrometastases in relation to the sectioning interval and the number of sections examined. METHODS: The authors examined 109 patients with micrometastatic SLNs from a series of 634 patients with carcinoma of the breast who underwent SLN biopsy and complete ALND as part of the surgical treatment for their disease. The SLNs were sectioned completely at 50-microm intervals, and the sections were examined intraoperatively. RESULTS: The overall frequency of metastases in axillary non-SLNs was 21.8%. The frequency was correlated significantly with the size of the SLN micrometastatic focus (P = 0.02): 36.4% of patients with foci > 1 mm had metastases in axillary lymph nodes--a percentage approaching 44.7% of patients with macrometastatic SLNs--whereas only 15.6% of patients with micrometastases < or = 1 mm had other involved axillary lymph nodes. CONCLUSIONS: Outside of clinical trials, patients with T1 and small T2 breast carcinoma and micrometastatic SLNs should undergo complete ALND for adequate staging. However, patients with SLN micrometastases up to 1 mm in greatest dimension have a significantly lower risk of additional axillary metastases, raising the question of whether ALND may be avoided in this subgroup of patients.     

Determinants of prostate cancer-specific survival after radiation therapy for patients with clinically localized prostate cancer.

PURPOSE: Identifying pretreatment and posttreatment predictors of time to prostate cancer-specific death (PCSD) after external-beam radiation therapy (RT) was the subject of this study. PATIENTS AND METHODS: A Cox regression analysis was used to evaluate the ability of the pretreatment risk group to predict time to PCSD for 381 patients who underwent RT for clinically localized prostate cancer. Posttreatment factors analyzed for the 94 patients who experienced prostate-specific antigen (PSA) failure included the time to PSA failure, the posttreatment PSA doubling time (DT), and the timing of salvage hormonal therapy. RESULTS: Despite the median age of 73 years at diagnosis, 45% of patients with high-risk disease were estimated to die from prostate cancer within 10 years after RT compared with 0% (P =.004) and 6% (P =.05) for patients with low- or intermediate-risk disease, respectively. Predictors of time to PCSD after PSA failure included PSA DT (P =.01) and delayed use of hormonal therapy (P 相似文献   

乳腺癌前哨淋巴结微小转移的检测

目的探讨检测乳腺癌前哨淋巴结(SLN)微小转移的最佳方法,研究临床病理因素与微小转移的相关性。方法应用同位素法检测乳腺癌SLN;对常规病理检查阴性的SLN,以100μm为间隔,进行多层间隔连续切片,并做HE和免疫组化染色检测微小转移;取肿瘤标本进行连续切片,并行免疫组化染色。结果共检测59例患者的121枚SLN和44份肿瘤标本,有14例(23.7%)患者的17枚(14.O%)SLN有微小转移。用HE染色法,切片数量从1层增加到3层时,微小转移的检出例数分别为3、7和10例;在3个层面上行间隔连续切片,HE分别与AE1/3、CK19和muc1联合检测时,微小转移的检出例数分别为14、12和16例。增加切片数量或采用联合检测的方法,可以提高微小转移的检出数量,微小转移与原发肿瘤大小、c-erbB2、MMP-2和血管内皮生长因子(VEGF)的表达相关。结论检测SLN微小转移的最佳方法为间隔100μm、在2个层面上行间隔连续切片,同时进行HE和muc1染色,可以检出绝大多数的微小转移。